AIM: To investigate the effect of granulocyte colony stimulating factor (G-CSF) on arabinosyl cytosine (Ara-C)-induced apoptosis in HL-60 leukemia cells. METHODS: The proliferation of HL-60 leukemia cell was observed by hemopoietic cell culture. Apoptosis was measured by the morphology of apoptosis cell , the quantitation of DNA fragmentation with the diphenylamine reaction. The change in drug sensitivity was measured by “MTT”. RESULTS: G-CSF could stimulate the proliferation of HL-60 leukemia cell and colonies of cell increased to 76.5?18.0, compared to the control group (46.5?13.5. P

Objective To investigate the effect of construct the Notch1 (NICD) eukaryotic expression vector on the proliferation and differentiation of rat bone marrow mesenchymal stem cells (BMSCs) in vitro. Methods Rat BMSCs were experimented as the object. NICD eukaryotic expression vector was constructed. pEGFP-N1-NICD expressing plasmids were used to transfect BMSCs. The study included control group (CON group), empty vector group (VEC group) and the trans-fection group (TRA group). After 48-hour transfection, BMSCs were observed for general morphology. The protein expres-sions of NSE, GFAP and Notch1 were detected by real-time PCR and Western blotting assay respectively. The apoptosis, cy-cle distribution and cell proliferation were evaluated by flow cytometry and MTT assay. Results The DNA sequencing con-firmed that the pEGFP-N1-NICD recombinant plasmid was successfully constructed, and both VEC group and TRA group expressed green fluorescence after 48-hour transfection. The relative expression levels of Notch1 and GFAP mRNA and pro-tein were significantly higher in TRA group than those in VEC group and CON group (P<0.05), and there was no significant difference between VEC group and CON group. After 48-hour transfection, the ratio of living cells was significantly lower in TRA group than that of CON group and VEC group, and the early apoptotic rate and late apoptotic rate were significantly higher in TRA group than those of CON group and VEC group (P<0.05). The late apoptotic rate was significantly higher in VEC group than that of CON group. The proportion of G1/G0 cells was significantly higher in TRA group than that of CON group and VEC group, but S and G2/M cells were significantly lower (P<0.05). The value of growth curve was gradually de-creased in TRA group than that of CON group and VEC group (P<0.05). Conclusion The high expression of NICD gene might induce apoptosis of BMSCs, inhibit the proliferation in part, and induce into glial-like cell differentiation.

Objective To analyze results of atopy patch test (APT) with dust mite allergens at different concentrations in patients with atopic dermatitis (AD),and to evaluate the diagnostic value of APT.Methods Totally,85 patients with AD were enrolled into this study.All the patients underwent APT with 5 concentrations (3 000,5 000,7 000,10 000 and 12 000 pnu/g) of dust mite allergens,as well as skin prick test (SPT) with dust mite allergens.Dust mite allergens were obtained from two different manufacturers (group 1 and 2).Enzyme-linked immunosorbent assay (ELISA) was performed to detect allergen-specific immunoglobulin E (SIgE) in sera from these patients.The sensitivity,specificity and positive predictive value of APT,SPT and SIgE assay were compared,and the results of APT were compared among different concentrations of allergens and between allergens from different manufacturers.Results When SIgE assay served as the gold standard,the sensitivity,specificity and positive predictive value in the diagnosis of dust mite allergy were 79.41％,76.12％,and 64.29％ respectively for APT,73.53％,80.95％ and 67.57％ respectively for SPT with group 1 dust mite allergens,and 81.53％,77.78％ and 65.09％ respectively for APT,76.02％,79.85％ and 66.07％ respectively for SPT with group 2 allergens.When SPT was regarded as the gold standard,the sensitivity,specificity and positive predictive value in the diagnosis of dust mite allergy were 78.38％,77.42％,67.44％ respectively for APT,67.57％,84.21％,73.53％ respectively for SIgE assay with group 1 dust mite allergens,79.25％,80.63％ and 69.55％ respectively for APT,61.07％,82.54％ and 77.21％ respectively for SIgE assay with group 2 allergens.There were no significant differences in the sensitivity,specificity or positive predictive value of APT or SPT between the two groups of allergens.The positive rate of APT was 8.24％,22.35％,29.41％,44.71％ and 41.18％ respectively with group 1 allergens at 3 000,5 000,7 000,10 000 and 12 000 pnu/g,and 3.53％,23.53％,31.76％,34.12％ and 35.29％ respectively with group 2 allergens.No significant differences were observed in the positive rate of APT between group 1 and 2 allergens at same concentrations (all P ＞ 0.05),but a significant difference was observed in that between different concentrations of group 1 or 2 allergens (both P＜ 0.05).The positive rate of APT increased with the increase of allergen concentrations,but stopped rising when the concentrations of group 1 and 2 allergens reached 7 000 pnu/g and 5 000 pnu/g respectively.Conclusions The sensitivity of APT is relatively high for the diagnosis of dust mite allergy.The positive rate of APT increased with the increase in allergen concentrations,but stopped rising when the concentrations reached a certain level.

Objective To elucidate the mutations of NPHS1 gene in children with sporadic steroid-resistant nephrotic syndrome (SRNS) in Southern Chinese Han ethnic group.Methods Peripheral blood samples were collected for genetic analysis from 40 patients with sporadic SRNS and 50 healthy volunteers as control.Genomic DNA was isolated from peripheral blood leucocytes.Twenty-nine exons and exon-intron boundaries of the NPHS1 gene were amplified by polymerase chain reaction.Mutational analysis was performed by DNA sequencing directly.Results Seven variants,928G＞A(D310N),2677A＞G (T893A),2869G＞C (V957L),IVS8+30C＞T,IVS21+14G＞A,IVS25-23C＞T and *142T＞C,of NPHS1 gene were found in 6 of 40 children with sporadic SRNS,whereas they were not found in 50 healthy controls.2677A ＞G,IVS8 +30C ＞T,IVS21 +14G＞A,IVS25-23C ＞T and *142T＞C were novel.Moreover,thirteen already reported NPHS1 polymorphisms,294C＞T,349G＞A,IVS3+15C＞T,IVS3+61A＞G,803G＞A,IVS8+68A＞G,1339G ＞A,1802G ＞C,2223C ＞T,2289C ＞T,IVS24 +36C ＞T,3315G＞A and IVS27 +45C ＞T,were detected in some patients and controls. Conclusions NPHS1 mutations in 6 of 40 children with sporadic SRNS in Southern Chinese Han ethnic group (15％) are detected.NPHS1 mutations are existed in Southern Chinese children,so it is necessary to perform the mutation analysis of NPHS1 gene in those children patients.

OBJECTIVE:To optimize the ultrasonic extraction technology for Tianshu capsules. METHODS:Using the transfer rate of active ingredient ferulic acid in Ligusticum chuanxiong and gastrodin in Gastrodia elata of Tianshu capsules as investigation indexes,L9(34)orthogonal design test was adopted to investigate the effects of ethanol volume fraction,ethanol amount,extraction time and ultrasonic power on the extraction rate. Ultrasonic extraction technology for Tianshu capsules was optimized,and verifica-tion test was carried out. RESULTS:The optimized extraction technology for Tianshu capsules was as follow as 8-fold 70% etha-nol,extracting twice under 350 W,extracting 1.5 h every time. Results of verification test showed the average transfer rate was 94.06% for ferulic acid(RSD=0.18%,n=3)and 95.02% for gastrodin(0.47%,n=3). CONCLUSIONS:The optimized tech-nology is rapid,simple,stable and feasible,and can be used for extracting the active ingredients in Tianshu capsules.

OBJECTIVETo investigate the statement on randomized controlled trials on Wenxin granule for treatment of atrial fibrillation and to judge whether those trials could offer high quality evidence or not, thus improve design level and quality.

METHODRCTs were searched from home and abroad about atrial fibrillation treated with Wenxin granule, which reported before October, 2010. Jadad scale and CONSORT statement were used.

RESULTThere were 66 RCTs retrieved that met inclusion criteria. Using Jadad rating scale, only 2 literatures gain score 4 and 1 literature gains score 3, 54 literatures gain score 2, 7 literatures gain score 1, 2 literatures gain score 0. Only 2 literatures described random number table as the method of grouping. None of the RCTs was reported the allocation concealment. Only 1 literature was used blinding. Fifty-nine literatures were mentioned the lost to follow-up conditions. According to the CONSORT standards, only six literatures (9.1%) mentioned the method of generating the random sequence. Four literatures (6.1%) were quasi-random. Nineteen literatures (28.8%) had inclusion criteria. Six literatures (9.1%) had the follow-up record. Fifty one literatures (77.3%) described the adverse events. None had the estimation of the sample size, intention-to-treat analysis and stratified analysis. None had the ethical approval or informed consent.

CONCLUSIONThe quality of clinical trials of Wenxin granule in treating atrial fibrillation needs to be improved.

Objective:To study the correlation between hearing loss and cognitive decline in community-dwelling older adults and to analyze the influencing factors.Methods:Using Pure-tone audiometry, the Hearing Handicap Inventory for the Elderly (HHIE), Mini-Mental State Examination (MMSE), Patient Health Questionnaire-9 (PHQ-9), and Generalized Anxiety Disorder, a survey was conducted with 492 community-dwelling older adults aged 60 years and above in Beijing. Age, gender, education level, hearing loss, levels of anxiety and depression, and other factors were used as independent variables, and the MMSE scores were used as dependent variables. Analysis of variance and multiple linear regression were performed.Results:Among the 492 subjects, based on the better ears, 418 (85.0%) had hearing loss and 160 (32.5%) had disabled hearing loss. Factors such as age (β=-0.33, P<0.01), gender (β=0.09, P=0.04), education level (β=0.31, P<0.01), hearing loss (β=-0.11, P=0.02) and marital status (β=0.56, P=0.02) were significantly related to cognitive decline. Gender (β=-0.17, P<0.01) and hearing loss (β=0.08, P=0.02) correlated with anxiety indicators; gender (β=-0.84, P=0.02) and hearing loss (β=0.04, P=0.01) also correlated with depression indicators. Conclusion:Results indicated a correlation between hearing loss and cognitive decline, and between hearing loss and anxiety and depression among community-dwelling older adults in Beijing.